Novel Thienopyrimidones

ABSTRACT

This invention relates to thienopyrimidinones and their use as inhibitors of TRPA1 activity, pharmaceutical compositions containing the same, and methods of using the same as agents for treatment and/or prevention of fibrotic diseases, inflammatory and auto-immune diseases and CNS-related diseases.

FIELD OF THE INVENTION

This invention relates to thienopyrimidinones and their use asinhibitors of TRPA1 activity, pharmaceutical compositions containing thesame, and methods of using the same as agents for treatment and/orprevention of fibrotic diseases, inflammatory and auto-immune diseasesand CNS-related diseases.

BACKGROUND INFORMATION

Transient receptor potential channels (TRP channels) are a group ofvoltage-gated ion channels located mostly on the plasma membrane ofnumerous mammalian cell types. There are approximately 30 structurallyrelated TRP channels sorted into groups: TRPA, TRPC, TRPM, TRPML, TRPN,TRPP and TRPV. Transient receptor potential cation channel, subfamily A,member 1 (TRPA1), also known as transient receptor potential ankyrin 1,is the only member of the TRPA gene subfamily Structurally, TRPAchannels are characterized by multiple N-terminal ankyrin repeats (˜14in the N-terminus of human TRPA1) that gives rise to the “A” for ankyrindesignation (Montell, 2005).

TRPA1 is highly expressed in the plasma membrane of sensory neurons inthe dorsal root and nodose ganglia that serve both skin and lung, aswell as in small intestine, colon, pancreas, skeletal muscle, heart,brain, bladder and lymphocytes (https://www.proteinatlas.org/) as wellas in human lung fibroblasts.

TRPA1 is best known as a sensor for environmental irritants giving riseto somatosensory modalities such as pain, cold and itch. TRPA1 isactivated by a number of reactive, electrophilic stimuli (e.g. allylisothiocyanate, reactive oxygen species), as well as non-reactivecompounds (e.g. icilin), implicated in cough associated with asthma,chronic pulmonary obstructive disease (COPD), idiopathic pulmonaryfibrosis (IPF) or post-viral cough or for chronic idiopathic cough aswell as cough in sensitive patients. (Song and Chang, 2015; Grace andBelvisi, 2011). TRPA1 inhibitors are useful in the treatment of IPF inwhich cough is highly prevalent because of the link between cough andlung injury, based on studies showing cough-induced elevation of TGF-β(Xie et al., 2009; Froese et al., 2016; Tschumperlin et al., 2003;Yamamoto et al., 2002; Ahamed et al., 2008). TRPA1 antagonists inhibitcalcium signaling triggered by cough triggers such as cigarette smokeextract (CSE) oxidative stress, inflammatory mediator release anddownregulated antioxidant gene expression (Lin et al., 2015; Wang etal., 2019). TRPA1 antagonists are effective in studies of atopicdermatitis (Oh et al., 2013; Wilson et al., 2013), contact dermatitis(Liu et al., 2013), psoriasis-associated itch (Wilson et al., 2013) andIL-31-dependent itch (Cevikbas et al., 2014). A human TRPA1gain-of-function has been associated with familial episodic painsyndrome (Kremeyer et al., 2010). A TRPA1 antagonist was effective in abehavioral model of migraine-related allodynia (Edelmayer et al., 2012).TRPA1 is selectively increased in trigeminal ganglia innervating injuredteeth when compared to TRPA1 expression in trigeminal gangliainnervating healthy teeth (Haas et al., 2011). Several anaesthetics areknown to be TRPA1 agonists, including isoflurane (Matta et al., 2008)providing rationale for TRPA1 inhibitors for the relief of post-surgicalpain. TRPA1 knockout mice and wild type mice treated with a TRPA1antagonist showed anxiolytic- and antidepressant-like phenotypes (deMoura et al., 2014). TRPA1 inhibitors are expected to have benefit inthe treatment of diabetic neuropathy based on studies showing amechanistic link of inverse regulation between AMPK and TRPA1 (Hiyama etal., 2018; Koivisto and Pertovaara, 2013; Wang et al., 2018). TRPA1knockout mice exhibit smaller myocardial infarct sizes compared to wildtype mice (Conklin et al., 2019). TRPA1 knockout and pharmacologicalintervention inhibited TNBS-induced colitis in mice (Engel et al.,2011). In a mouse brain ischaemia model, TRPA1 knock-out and TRPA1antagonists reduce myelin damage (Hamilton et al., 2016). Urate crystalsand joint inflammation are reduced in TRPA1 knockout mice in amonosodium urate mouse model of gout (Moilanen et al., 2015). TRPA1deletion in rats ameliorated joint inflammation and hyperalgesia in arat model of acute gout flares (Trevisan et al., 2014). Activation ofTRPA1 elicits an inflammatory response in osteoarthritic chondrocytes(Nummenmaa et al., 2016). TRPA1 inhibition and genetic deletion reducesinflammatory mediators in osteoarthritic mouse chondrocytes and murinecartilage (Nummenmaa et al., 2016). Finally, TRPA1 knockout miceexhibited improvements in weight bearing on the osteoarthritic limb inan MIA-evoked knee swelling model (Horvath et al., 2016). TRPA1 isdifferentially expressed in the in the bladder epithelium rats (Du etal., 2007) and of patients with bladder outlet obstruction (Du et al.,2008). TRPA1 receptor modulation attenuates bladder overactivity in arat model of spinal cord injury (Andrade et al., 2011) and intrathecaladministration of TRPA1 antagonists attenuate cyclophosphamide-inducedcystitis in rats with hyper-reflexia micturition (Chen et al., 2016).

It is therefore desirable to provide potent TRPA1 inhibitors.

TRPA1 inhibitors of various structural classes are reviewed in S.Skerratt, Progress in Medicinal Chemistry, 2017, Volume 56, 81-115 andin D. Preti, G. Saponaro, A. Szallasi, Pharm. Pat. Anal. (2015) 4 (2),75-94.

WO2017/060488 discloses compounds that are antagonists of TRPA1, havingthe generalized structural formula

The TRPA1 activity of Examples 28 and 29 bearing a tetrazolyl ringtherein is not disclosed.

L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809 disclosesquinazolinone-based TRPA1 antagonists including compounds of thegeneralized structural formula

of which compound 31, wherein R is OH, is disclosed as having anantagonistic TRPA1 activity of IC₅₀ 58 nM in a FLIPR assay and having anintrinsic clearance in human liver microsomes of <14 μL/min/kg.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel thienopyrimidinones that aresurprisingly potent inhibitors of TRPA1 (Assay A), further characterisedby

-   -   improved stability in human liver microsomes (Assay B)    -   improved stability in human hepatocytes (Assay C).

Compounds of the present invention differ structurally from examples 28and 29 in WO2017/060488 in that they contain a thienopyrimidinone corewith amido substituents as well as substituents adjacent to a secondaryaliphatic alcohol. Compounds of the present invention additionallydiffer structurally from example 31 in L. Schenkel, et al., J. Med.Chem. 2016, 59, 2794-2809, in that they bear a tetrazolyl ring. Thesestructural differences unexpectedly lead to a favourable combination of(i) inhibition of TRPA1, (ii) stability in human liver microsomes, (iii)stability in human hepatocytes, and (iv) fewer unwanted humanmetabolites.

Compounds of the invention are thus superior to those disclosed in theprior art in terms of the combination of the following parameters:

-   -   potency as inhibitors of TRPA1    -   stability in human liver microsomes    -   stability in human hepatocytes    -   giving rise to fewer unwanted human metabolites

Stability in human liver microsomes refers to the susceptibility ofcompounds to biotransformation in the context of selecting and/ordesigning drugs with favorable pharmacokinetic properties as a firstscreening step. The primary site of metabolism for many drugs is theliver. Human liver microsomes contain the cytochrome P450s (CYPs), andthus represent a model system for studying phase I drug metabolism invitro Enhanced stability in human liver microsomes is associated withseveral advantages, including increased bioavailability and adequatehalf-life, which can enable lower and less frequent dosing of patients.Thus, enhanced stability in human liver microsomes is a favorablecharacteristic for compounds that are to be used for drugs. Therefore,compounds of the present invention in addition to being able to inhibitTRPA1 are expected to have a favorable in vivo clearance and thus thedesired duration of action in humans.

Stability in human hepatocytes refers to the susceptibility of compoundsto biotransformation in the context of selecting and/or designing drugswith favorable pharmacokinetic properties. The primary site ofmetabolism for many drugs is the liver. Human hepatocytes contain thecytochrome P450s (CYPs) and other drug metabolizing enzymes, and thusreps resent a model system for studying drug metabolism in vitro.(Importantly, in contrast to liver microsomes assay, the hepatocytesassay covers also phase II biotransformations as well as liver-specifictransporter-mediated processes, and therefore represents a more completesystem for drug metabolism studies) Enhanced stability in humanhepatocytes is associated with several advantages, including increasedbioavailability and adequate half-life, which can enable lower and lessfrequent dosing of patients. Thus, enhanced stability in humanhepatocytes is a favorable characteristic for compounds that are to beused for drugs.

The present invention provides novel compounds according to formula (I)

wherein

A is selected from the group consisting of phenyl, naphthyl, thiophenyl,benzothiophenyl or benzofuranyl, optionally substituted with one or twomembers of the group consisting of H, F, Cl, Br, C₁₋₄-alkyl,F₁₋₃-fluoro-C₁₋₄-alkyl, CN, OCH₃, cyclopropyl, and cyclobutyl,

or

A is selected from

and

R¹ is selected from H, C₁₋₄-alkyl, F₁₋₃-fluoro-C₁₋₄-alkyl, C₁₋₄-alkyl-OHor C₁₋₄-alkyl-CN;

R² is selected from C₁₋₂-alkyl or Cl;

or R¹ and R² are each CH₂ joined via a bond forming a 6-membered ring;

R³ is selected from H, or C₁₋₄-alkyl.

Another embodiment of the present invention relates to a compound offormula (I), wherein A is selected from the group consisting of phenyl,thiophenyl or benzofuranyl, optionally substituted with one or twomembers of the group consisting of H, F, Cl, Br, C₁₋₄-alkyl,F₁₋₃-fluoro-C₁₋₄-alkyl, CN, OCH₃, cyclopropyl, and cyclobutyl,

or

A is selected from

and substituents R¹ and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein A is selected from the group consisting of phenyl,thiophenyl or benzofuranyl, optionally substituted with one or twomembers of the group consisting of H, F, Cl, Br, C₁₋₄-alkyl,F₁₋₃-fluoro-C₁₋₄-alkyl, CN, OCH₃, cyclopropyl, and cyclobutyl,

or

A is selected from

and substituents R¹ and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein A is selected from the group consisting of phenyl,thiophenyl or benzofuranyl, optionally substituted with one or twomembers of the group consisting of H, F, Br, Cl, and CH₃,

or

A is selected from

and substituents R¹ and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein A is selected from the group consisting of phenyl,thiophenyl or benzofuranyl, optionally substituted with one or twomembers of the group consisting of H, F, Br, Cl, and —CH₃,

or

A is selected from

and substituents R¹ and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein A is selected from the group consisting of

and substituents R¹ and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein R¹ is selected from the group consisting of H andC₁₋₄-alkyl; and substituents A and R² are defined as in any of thepreceding embodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein R¹ is selected from the group consisting of H andH₃C; and substituents A and R² are defined as in any of the precedingembodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein R² is H₃C or Cl; and substituents A and R¹ aredefined as in any of the preceding embodiments.

Another embodiment of the present invention relates to a compound offormula (I), wherein R³ is H or H₃C; and substituents A, R¹ and R² aredefined as in any of the preceding embodiments.

Preferred is a compound of formula (I), selected from the groupconsisting of

and substituent A is defined as in any of the preceding embodiments.

Particularly preferred is the compound according to formula (I) selectedfrom the group consisting of

Used Terms and Definitions

Terms not specifically defined herein should be given the meanings thatwould be given to them by one of skill in the art in light of thedisclosure and the context. As used in the specification, however,unless specified to the contrary, the following terms have the meaningindicated and the following conventions are adhered to.

In the groups, radicals, or moieties defined below, the number of carbonatoms is often specified preceding the group, for example, C₁₋₆-alkylmeans an alkyl group or radical having 1 to 6 carbon atoms. In generalin groups like HO, H₂N, (O)S, (O)₂S, NC (cyano), HOOC, F₃C or the like,the skilled artisan can see the radical attachment point(s) to themolecule from the free valences of the group itself. For combined groupscomprising two or more subgroups, the last named subgroup is the radicalattachment point, for example, the substituent “aryl-C₁₋₃-alkyl” meansan aryl group which is bound to a C₁₋₃-alkyl-group, the latter of whichis bound to the core or to the group to which the substituent isattached.

In case a compound of the present invention is depicted in form of achemical name and as a formula in case of any discrepancy the formulashall prevail. An asterisk is may be used in sub-formulas to indicatethe bond which is connected to the core molecule as defined.

The numeration of the atoms of a substituent starts with the atom thatis closest to the core or to the group to which the substituent isattached.

For example, the term “3-carboxypropyl-group” represents the followingsubstituent:

wherein the carboxy group is attached to the third carbon atom of thepropyl group. The terms “1-methylpropyl-”, “2,2-dimethylpropyl-” or“cyclopropylmethyl-” group represent the following groups:

The asterisk may be used in sub-formulas to indicate the bond that isconnected to the core molecule as defined.

The term “C_(1-n)-alkyl”, wherein n is an integer selected from 2, 3, 4or 5, either alone or in combination with another radical denotes anacyclic, saturated, branched or linear hydrocarbon radical with 1 to n Catoms. For example the term C₁₋₅-alkyl embraces the radicals H₃C—,H₃C—CH₂—, H₃C—CH₂—CH₂—, H₃C—CH(CH₃)—, H₃C—CH₂—CH₂—CH₂—,H₃C—CH₂—CH(CH₃)—, H₃C—CH(CH₃)—CH₂—, H₃C—C(CH₃)₂—, H₃C—CH₂—CH₂—CH₂—CH₂—,H₃C—CH₂—CH₂—CH(CH₃)—, H₃C—CH₂—CH(CH₃)—CH₂—, H₃C—CH(CH₃)—CH₂—CH₂—,H₃C—CH₂—C(CH₃)₂—, H₃C—C(CH₃)₂—CH₂—, H₃C—CH(CH₃)—CH(CH₃)— andH₃C—CH₂—CH(CH₂CH₃)—.

The term “F_(1-m)-fluoro-C_(1-n)-alkyl”, wherein m is an integerselected from 2 or 3, and n is an integer selected from 2, 3, 4 or 5,denotes an C_(1-n)-alkyl group as defined hereinbefore wherein one ormore hydrogen atoms are replaced by 1-m fluorine. For example,F₁₋₃-fluoro-C₁₋₂-alkyl embraces the radicals FH₂C, F₂HC, F₃C, FH₂C—H₂C,F₂HC—H₂C, F₃CH₂C, FH₂C—FHC, FH₂C—F₂C, F₂HC—FHC, H₃C—FHC, and H₃C—F₂C.

The term phenyl refers to the radical of the following ring

The term naphthyl refers to the radical of the following ring

The term thiophenyl refers to the radical of the following ring

The term thienopyrimidone refers to the radical of the following ring

and includes

The term benzothiophenyl refers to the radical of the following ring

The term benzofuranyl refers to the radical of the following ring

The term cyclopropyl refers to the radical of the following ring

The term cyclobutyl refers to the radical of the following ring

The term tetrazolyl refers to the radical of the following ring

The term “substituted” as used herein, means that any one or morehydrogens on the designated atom is replaced with a selection from theindicated group, provided that the designated atom's normal valence isnot exceeded, and that the substitution results in a stable compound.

Unless specifically indicated, throughout the specification and theappended claims, a given chemical formula or name shall encompasstautomers and all stereo, optical and geometrical isomers (e.g.enantiomers, diastereomers, E/Z isomers etc.) and racemates thereof aswell as mixtures in different proportions of the separate enantiomers,mixtures of diastereomers, or mixtures of any of the foregoing formswhere such isomers and enantiomers exist, as well as salts, includingpharmaceutically acceptable salts thereof and solvates thereof such asfor instance hydrates including solvates of the free compounds orsolvates of a salt of the compound.

In general, substantially pure stereoisomers can be obtained accordingto synthetic principles known to a person skilled in the field, e.g. byseparation of corresponding mixtures, by using stereochemically purestarting materials and/or by stereoselective synthesis. It is known inthe art how to prepare optically active forms, such as by resolution ofracemic forms or by synthesis, e.g. starting from optically activestarting materials and/or by using chiral reagents.

Enantiomerically pure compounds of this invention or intermediates maybe prepared via asymmetric synthesis, for example by preparation andsubsequent separation of appropriate diastereomeric compounds orintermediates which can be separated by known methods (e.g. bychromatographic separation or crystallization) and/or by using chiralreagents, such as chiral starting materials, chiral catalysts or chiralauxiliaries.

Further, it is known to the person skilled in the art how to prepareenantiomerically pure compounds from the corresponding racemic mixtures,such as by chromatographic separation of the corresponding racemicmixtures on chiral stationary phases; or by resolution of a racemicmixture using an appropriate resolving agent, e.g. by means ofdiastereomeric salt formation of the racemic compound with opticallyactive acids or bases, subsequent resolution of the salts and release ofthe desired compound from the salt; or by derivatization of thecorresponding racemic compounds with optically active chiral auxiliaryreagents, subsequent diastereomer separation and removal of the chiralauxiliary group; or by kinetic resolution of a racemate (e.g. byenzymatic resolution); by enantioselective crystallization from aconglomerate of enantiomorphous crystals under suitable conditions; orby (fractional) crystallization from a suitable solvent in the presenceof an optically active chiral auxiliary.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use withoutexcessive toxicity, irritation, allergic response, or other problem orcomplication, and commensurate with a reasonable benefit/risk ratio. Asused herein, “pharmaceutically acceptable salt” refers to derivatives ofthe disclosed compounds wherein the parent compound forms a salt or acomplex with an acid or a base. Examples of acids forming apharmaceutically acceptable salt with a parent compound containing abasic moiety include mineral or organic acids such as benzenesulfonicacid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid,gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malicacid, malonic acid, mandelic acid, methanesulfonic acid,4-methyl-benzenesulfonic acid, phosphoric acid, salicylic acid, succinicacid, sulfuric acid and tartaric acid.

Examples for cations and bases forming a pharmaceutically acceptablesalt with a parent compound containing an acidic moiety include Na⁺, K⁺,Ca²⁺, Mg²⁺, NH₄ ⁺, L-arginine, 2,2′-iminobisethanol, L-lysine,N-methyl-D-glucamine or tris(hydroxymethyl)-aminomethane. Thepharmaceutically acceptable salts of the present invention can besynthesized from the parent compound that contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha sufficient amount of the appropriate base or acid in water or in anorganic diluent like ether, ethyl acetate, ethanol, isopropanol, oracetonitrile, or a mixture thereof.

Salts of other acids than those mentioned above which for example areuseful for purifying or isolating the compounds of the present invention(e.g. trifluoroacetate salts,) also comprise a part of the presentinvention.

Biological Assays

Assay A: TRPA1 Assay

The activity of the compounds of the invention may be demonstrated usingthe following in vitro TRPA1 cell assay:

Method:

A human HEK293 cell line over-expressing the human TRPA1 ion channel(Perkin Elmer, Product No. AX-004-PCL) was used as a test system forcompound efficacy and potency. Compound activity was determined bymeasuring the effect of compounds on intracellular calcium concentrationinduced by AITC (Allylisothiocyanat) agonism in a FLIPRtetra system(Molecular Devices).

Cell Culture:

The cells were obtained as frozen cells in cryo-vials and stored untiluse at −150° C. Cells were grown in culture medium (MEM/EBSS medium with10% FCS and 0.4 mg/ML Geneticin). It is important that density does notexceed 90% confluence. For sub-culturing cells were detached from flasksby Versene. At the day before the assay, cells were detached, washedtwice with medium (MEM/EBSS medium with 10% FCS) and 20000 cells in 20μl/well were seeded to Poly D-Lysin biocoated 384-well plates (black,clear bottom, Cat. 356697) from Corning. Plates were incubated for 24hours at 37° C./5% CO2 before use in the assay.

Compound Preparation

The test compounds were dissolved in 100% DMSO at a concentration of 10mM and in a first step diluted in DMSO to a concentration of 5 mM,followed by serial dilution steps in 100% DMSO. Dilution factor andnumber of dilution steps may vary according to needs. Typically 8different concentrations by 1:5 dilutions were prepared, furtherintermediate dilutions (1:20) of the substances were carried out withHBSS/HEPES buffer (1×HEPES, Cat. 14065 from Gibco, 20 mM HEPES, Cat.83264 from SIGMA, 0.1% BSA Cat. 11926 from Invitrogen, pH 7.4

FLIPR Assay:

At the assay day cells were washed 3× with assay puffer, 20 μL bufferremained in the wells after washing. 10 μL Ca6 kit (Cat. R8191MolecularDevices) loading buffer in HBSS/HEPES was added to the cellsand the plates were incubated with lid for 120 minutes at 37°/5% CO2. 10μL of compound or controls in HBSS/HEPES buffer/5% DMSO from theintermediate dilution plate were carefully added to the wellsLuminescence (indicating the calcium influx or release) was read on theFLIPRtetra device for 10 minutes to monitor the compound induced effects(e.g. agonism). Finally 10 μL of the agonist AITC 50 μM dissolved inHBSS/HEPES buffer/0.05% DMSO (final concentration 10 μM) was added tothe wells followed by an additional read on the FLIPRtetra device for 10minutes. The area under the signal curve (AUC) after AITC addition wasused for IC50/% inhibition calculations

Data Evaluation and Calculation:

Each assay microtiter plate contained wells with vehicle (1% DMSO)controls instead of compound as controls for AITC induced luminescence(100% CTL; high controls) and wells with vehicle controls without AITCas controls for non-specific changes in luminescence (0% CTL; lowcontrols).

The analysis of the data was performed by the calculation of the areaunder signal curve of the individual wells. Based on this values the %value for the measurement of each subs stance concentration wascalculated (AUC(sample)−AUC(low))*100/(AUC(high)−AUC(low)) using MegaLabsoftware (in house development). The IC50 values were calculated fromthe % control values using MegaLab software. Calculation:[y=(a−d)/(1+(x/c){circumflex over ( )}b)+d], a=low value, d=high value;x=conc M; c=IC50 M; b=hill; y=% ctrl

TABLE 1 Biological data for compounds of the invention as obtained inAssay A hTRPA1 IC₅₀ Example [nM] 1 11 2 12 3 12 4 12 5 74 6 11 7 16 8 159 21 10 28 11 25 12 17 13 20 14 76 15 38 16 37 17 14 18 14 19 31 20 1421 80 22 7 23 6

TABLE 2 Biological data for prior art compounds (examples 28 and 29 inWO2017/060488) as obtained in Assay A. Example in hTRPA1 IC₅₀WO2017/060488 [nM] 28 366 29 1120

TABLE 3 Biological data for prior art compounds (example 31 in L.Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809) as obtained inAssay A. Example in Med. Chem. hTRPA1 IC₅₀ 2016, 59, 2794-2809 [nM] 3152

Assay B: Microsomal Clearance:

The metabolic degradation of the test compound is assayed at 37° C. withpooled liver microsomes. The final incubation volume of 100 μl per timepoint contains TRIS buffer pH 7.6 at RT (0.1 M), magnesium chloride (5mM), microsomal protein (1 mg/ml) and the test compound at a finalconcentration of 1 μM.

Following a short preincubation period at 37° C., the reactions areinitiated by addition of beta-nicotinamide adenine dinucleotidephosphate, reduced form (NADPH, 1 mM) and terminated by transferring analiquot into solvent after different time points (0, 5, 15, 30, 60 min).Additionally, the NADPH-independent degradation is monitored inincubations without NADPH, terminated at the last time point. The [%]remaining test compound after NADPH independent incubation is reflectedby the parameter c(control) (metabolic stability). The quenchedincubations are pelleted by centrifugation (10000 g, 5 min).

An aliquot of the supernatant is assayed by LC-MS/MS for the amount ofparent compound. The half-life (t½ INVITRO) is determined by the slopeof the semilogarithmic plot of the concentration-time profile.

The intrinsic clearance (CL_INTRINSIC) is calculated by considering theamount of protein in the incubation:

CL_INTRINSIC [μl/min/mg protein]=(Ln 2/(half-life [min]*protein content[mg/ml]))*1000

CL_INTRINSIC_INVIVO [ml/min/kg]=(CL_INTRINSIC [μL/min/mg protein]×MPPGL[mg protein/g liver]×liver factor [g/kg bodyweight])/1000

Qh [%]=CL [ml/min/kg]/hepatic blood flow [ml/min/kg])

Hepatocellularity, human: 120×10e6 cells/g liver

Liver factor, human: 25.7 g/kg bodyweight

Blood flow, human: 21 ml/(min×kg)

TABLE 4 Biological data for compounds of the invention as obtained inAssay B Example human LM [% Qh] 1 <23 2 <23 3 <23 4 <23 5 <23 6 <23 7 368 43 9 <23 10 <23 11 <23 12 <23 13 <23 14 <23 15 <23 16 <23 17 <23 18 3219 <23 20 <23 21 27 22 35 23 <23

TABLE 5 Biological data for prior art compounds (examples 28 and 29 inWO2017/060488) as obtained in Assay B. Example in human LM WO2017/060488[% Qh] 28 62 29 <23

TABLE 6 Biological data for prior art compounds (example 31 in L.Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809) as obtained inAssay B. Example in Med. Chem. human LM 2016, 59, 2794-2809 [% Qh] 31<23

Assay C: Hepatocyte Clearance

The metabolic degradation of the test compound is assayed in ahepatocyte suspension. Hepatocytes (cryopreserved) are incubated in.Dulbecco's modified eagle medium (supplemented with 3.5 μg glucagon/500mL, 2.5 mg insulin/500 mL and 3.75 mg/500 mL hydrocortison) containing5% or 50% species serum.

Following a 30 min preincubation in an incubator (37° C., 10% CO2) 5 μlof test compound solution (80 μM; from 2 mM in DMSO stock solutiondiluted 1:25 with medium) are added into 395 μl hepatocyte suspension(cell density in the range 0.25-5 Mio cells/mL depending on the species,typically 1 Mio cells/mL; final concentration of test compound 1 μM,final DMSO concentration 0.05%).

The cells are incubated for six hours (incubator, orbital shaker) andsamples (25μl) are taken at 0, 0.5, 1, 2, 4 and 6 hours. Samples aretransferred into acetonitrile and pelleted by centrifugation (5 min).The supernatant is transferred to a new 96-deepwell plate, evaporatedunder nitrogen and resuspended.

Decline of parent compound is analyzed by HPLC-MS/MS

CLint is calculated as followsCL_INTRINSIC=Dose/AUC=(CO/CD)/(AUD+clast/k)×1000/60. CO: initialconcentration in the incubation [μM], CD: cell density of vital cells[10e6 cells/mL], AUD: area under the data [μM×h], clast: concentrationof last data point [μM], k: slope of the regression line for parentdecline [h−1].

The calculated in vitro hepatic intrinsic clearance can be scaled up tothe intrinsic in vivo hepatic Clearance and used to predict hepatic invivo blood clearance (CL) by the use of a liver model (well stirredmodel).

CL_INTRINSIC_INVIVO [ml/min/kg]=(CL_INTRINSIC [μL/min/10e6cells]×hepatocellularity [10e6 cells/g liver]×liver factor [g/kgbodyweight])/1000

CL [ml/min/kg]=CL_INTRINSIC_INVIVO [ml/min/kg]×hepatic blood flow[ml/min/kg]/(CL_INTRINSIC_INVIVO [ml/min/kg]+hepatic blood flow[ml/min/kg])

Qh [%]=CL [ml/min/kg]/hepatic blood flow [ml/min/kg])

Hepatocellularity, human: 120×10e6 cells/g liver

Liver factor, human: 25.7 g/kg bodyweight

Blood flow, human: 21 ml/(min×kg)

TABLE 7 Biological data for compounds of the invention as obtained inAssay C human Hepatocytes Example [% Qh] 1 12 2 13 3 40 4 7 5 n.d. 6 7 726 8 36 9 16 10 14 11 28 12 31 13 9 14 13 15 18 16 17 17 22 18 48 19 2620 28 21 5 22 44 23 12

TABLE 8 Biological data for prior art compounds (examples 28 and 29 inWO2017/060488) as obtained in Assay C. Example in human HepatocytesWO2017/060488 [% Qh] 28 49 29 22

TABLE 9 Biological data for prior art compounds (example 31 in L.Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809) as obtained inAssay C. Example in Med. Chem. human Hepatocytes 2016, 59, 2794-2809 [%Qh] 31 73

Assay D: Metabolism in Human Hepatocytes In Vitro

The metabolic pathway of a test compound is investigated using primaryhuman hepatocytes in suspension. After recovery from cryopreservation,human hepatocytes are incubated in Dulbecco's modified eagle mediumcontaining 5% human serum and supplemented with 3.5 glucagon/500 ml, 2.5mg insulin/500 ml and 3.75 mg/500 ml hydrocortisone.

Following a 30 min preincubation in a cell culture incubator (37° C.,10% CO₂), test compound solution is spiked into the hepatocytesuspension to obtain a final cell density of 1.0*10⁶ to 4.0*10⁶ cells/ml(depending on the metabolic turnover rate of the compound observed withprimary human hepatocytes), a final test compound concentration of 10μM, and a final DMSO concentration of 0.05%.

The cells are incubated for six hours in a cell culture incubator on ahorizontal shaker, and samples are removed from the incubation after 0,0.5, 1, 2, 4 or 6 hours, depending on the metabolic turnover rate.Samples are quenched with acetonitrile and pelleted by centrifugation.The supernatant is transferred to a 96-deepwell plate, evaporated undernitrogen and resuspended prior to bioanalysis by liquidchromatography-high resolution mass spectrometry for identification ofputative metabolites.

TABLE 10 Major metabolites observed for Example 6 of the invention asobtained by Assay D: Cpd Metabolite M1 Mechanism Metabolite M1

Oxygenation Metabolite M2

Dehydrogenation Metabolite M3

Glucuronidation

The integrity of tetrazole ring was not affected in any of the majordescribed metabolites, based on tentative LC-MS^(n) structureelucidation.

Method of Treatment

The present invention is directed to compounds of general formula 1which are useful in the prevention and/or treatment of a disease and/orcondition associated with or modulated by TRPA1 activity, including butnot limited to the treatment and/or prevention of fibrotic disease,inflammatory and immunoregulatory disorders, respiratory orgastrointestinal diseases or complaints, ophthalmic diseases,inflammatory diseases of the joints and inflammatory diseases of thenasopharynx, eyes, and skin. Said disorders, diseases and complaintsinclude cough, idiopathic pulmonary fibrosis, other pulmonaryinterstitial diseases and other fibrotic, asthma or allergic diseases,eosinophilic diseases, chronic obstructive pulmonary disease, as well asautoimmune pathologies, such as rheumatoid arthritis andatherosclerosis, pain and neurological disorders, such as depression.

The compounds of general formula 1 are useful for the prevention and/ortreatment of: (1) Cough such as chronic idiopathic cough or chronicrefractory cough, cough associated with asthma, COPD and lung cancer andpost-viral cough.

(2) Pulmonary fibrotic diseases such as pneumonitis or interstitialpneumonitis associated with collagenosis, e.g. lupus erythematodes,systemic scleroderma, rheumatoid arthritis, polymyositis anddermatomysitis, idiopathic interstitial pneumonias, such as pulmonarylung fibrosis (IPF), non-specific interstitial pneumonia, respiratorybronchiolitis associated interstitial lung disease, desquamativeinterstitial pneumonia, cryptogenic orgainizing pneumonia, acuteinterstitial pneumonia and lymphocytic interstitial pneumonia,lymangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhan's cellhistiocytosis, pleural parenchymal fibroelastosis, interstitial lungdiseases of known cause, such as interstitial pneumonitis as a result ofoccupational exposures such as asbestosis, silicosis, miners lung (coaldust), farmers lung (hay and mould), Pidgeon fanciers lung (birds) orother occupational airbourne triggers such as metal dust ormycobacteria, or as a result of treatment such as radiation,methotrexate, amiodarone, nitrofurantoin or chemotherapeutics, or forgranulomatous disease, such as granulomatosis with polyangitis,Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, orinterstitial pneumonitis caused by different origins, e.g. aspiration,inhalation of toxic gases, vapors, bronchitis or pneumonitis orinterstitial pneumonitis caused by heart failure, X-rays, radiation,chemotherapy, M. boeck or sarcoidosis, granulomatosis, cystic fibrosisor mucoviscidosis, or alpha-1-antitrypsin deficiency.

(3) Other fibrotic diseases such as hepatic bridging fibrosis, livercirrhosis, non-alcoholic steatohepatitis (NASH), atrial fibrosis,endomyocardial fibrosis, old myocardial infarction, glial scar, arterialstiffness, arthrofibrosis, Dupuytren's contracture, keloid,scleroderma/systemic sclerosis, mediastinal fibrosis, myelofibrosis,Peyronie's disease, nephrogenic systemic fibrosis, retroperitonealfibrosis, adhesive capsulitis.

(4) Inflammatory, auto-immune or allergic diseases and conditions suchas allergic or non-allergic rhinitis or sinusitis, chronic sinusitis orrhinitis, nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis,asthma, pediatric asthma, allergic bronchitis, alveolitis, hyperreactiveairways, allergic conjunctivitis, bronchiectasis, adult respiratorydistress syndrome, bronchial and pulmonary edema, bronchitis orpneumonitis, eosinophilic cellulites (e.g., Well's syndrome),eosinophilic pneumonias (e.g., Loeffler's syndrome, chronic eosinophilicpneumonia), eosinophilic fasciitis (e.g., Shulman's syndrome),delayed-type hypersensitivity, non-allergic asthma; exercise inducedbronchoconstriction; chronic obstructive pulmonary disease (COPD), acutebronchitis, chronic bronchitis, cough, pulmonary emphysema; systemicanaphylaxis or hypersensitivity responses, drug allergies (e.g., topenicillin, cephalosporin), eosinophiliamyalgia syndrome due to theingestion of contaminated tryptophane, insect sting allergies;autoimmune diseases, such as rheumatoid arthritis, Graves' disease,Sjogren's syndrome psoriatic arthritis, multiple sclerosis, systemiclupus erythematosus, myasthenia gravis, immune thrombocytopenia (adultITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic anemia(auto-immune and drug induced), Evans syndrome (platelet and red cellimmune cytopaenias), Rh disease of the newborn, Goodpasture's syndrome(anti-GBM disease), Celiac, autoimmune cardio-myopathy juvenile onsetdiabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease;graft rejection (e.g., in transplantation), including allograftrejection or graft versus-host disease; inflammatory bowel diseases,such as Crohn's disease and ulcerative colitis; spondyloarthropathies;scleroderma; psoriasis (including T-cell mediated psoriasis) andinflammatory dermatoses such as an dermatitis, eczema, atopicdermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g.,necrotizing, cutaneous, and hypersensitivity vasculitis); erythemanodosum; eosinophilic myositis, eosinophilic fasciitis, cancers withleukocyte infiltration of the skin or organs; ophthalmic diseases suchas age related macular degeneration, diabetic retinopathy and diabeticmacular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis,vernal keratoconjunctivitis, scarring, anterior segment scarring,blepharitis, bless pharoconjunctivitis, bullous disorders, cicatricialpemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye,episcleritis, glaucoma, gliosis, Granuloma annulare, Graves'ophthalmopathy, intraocular melanoma, Pinguecula, proliferativevitreoretinopathy, pterygia, scleritis, uveitis, acute gout flares, goutor osteoarthritis.

(5) Pain such as chronic idiopathic pain syndrome, neuropathic pain,dysesthesia, allodynia, migraine, dental pain and post-surgical pain.

(6) Depression, anxiousness, diabetic neuropathy and bladder disorderssuch as bladder outlet obstruction, overactive bladder, cystitis;myocardial reperfusion injury or brain ischaemia injury.

Accordingly, the present invention relates to a compound of generalformula 1 for use as a medicament.

Furthermore, the present invention relates to the use of a compound ofgeneral formula 1 for the treatment and/or prevention of a diseaseand/or condition associated with or modulated by TRPA1 activity.

Furthermore, the present invention relates to the use of a compound ofgeneral formula 1 for the treatment and/or prevention of fibroticdisease, inflammatory and immunoregulatory disorders, respiratory orgastrointestinal diseases or complaints, ophthalmic diseases,inflammatory diseases of the joints and inflammatory diseases of thenasopharynx, eyes, and skin. Said disorders, diseases and complaintsinclude cough, idiopathic pulmonary fibrosis, other pulmonaryinterstitial diseases and other fibrotic, asthma or allergic diseases,eosinophilic diseases, chronic obstructive pulmonary disease, as well asautoimmune pathologies, such as rheumatoid arthritis andatherosclerosis.

Furthermore, the present invention relates to the use of a compound ofgeneral formula 1 for the treatment and/or prevention of:

(1) Cough such as chronic idiopathic cough or chronic refractory cough,cough associated with asthma, COPD and lung cancer and post-viral cough.

(2) Pulmonary fibrotic diseases such as pneumonitis or interstitialpneumonitis associated with collagenosis, e.g. lupus erythematodes,systemic scleroderma, rheumatoid arthritis, polymyositis anddermatomysitis, idiopathic interstitial pneumonias, such as pulmonarylung fibrosis (IPF), non-specific interstitial pneumonia, respiratorybronchiolitis associated interstitial lung disease, desquamativeinterstitial pneumonia, cryptogenic orgainizing pneumonia, acuteinterstitial pneumonia and lymphocytic interstitial pneumonia,lymangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhan's cellhistiocytosis, pleural parenchymal fibroelastosis, interstitial lungdiseases of known cause, such as interstitial pneumonitis as a result ofoccupational exposures such as asbestosis, silicosis, miners lung (coaldust), farmers lung (hay and mould), Pidgeon fanciers lung (birds) orother occupational airbourne triggers such as metal dust ormycobacteria, or as a result of treatment such as radiation,methotrexate, amiodarone, nitrofurantoin or chemotherapeutics, or forgranulomatous disease, such as granulomatosis with polyangitis,Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, orinterstitial pneumonitis caused by different origins, e.g. aspiration,inhalation of toxic gases, vapors, bronchitis or pneumonitis orinterstitial pneumonitis caused by heart failure, X-rays, radiation,chemotherapy, M. boeck or sarcoidosis, granulomatosis, cystic fibrosisor mucoviscidosis, or alpha-1-antitrypsin deficiency.

(3) Other fibrotic diseases such as heaptic bridging fibrosis, livercirrhosis, non-alcoholic steatohepatitis (NASH), atrial fibrosis,endomyocardial fibrosis, old myocardial infarction, glial scar, arterialstiffness, arthrofibrosis, Dupuytren's contracture, keloid,scleroderma/systemic sclerosis, mediastinal fibrosis, myelofibrosis,Peyronie's disease, nephrogenic systemic fibrosis, retroperitonealfibrosis, adhesive capsulitis.

(4) Inflammatory, auto-immune or allergic diseases and conditions suchas allergic or non-allergic rhinitis or sinusitis, chronic sinusitis orrhinitis, nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis,asthma, pediatric asthma, allergic bronchitis, alveolitis, hyperreactiveairways, allergic conjunctivitis, bronchiectasis, adult respiratorydistress syndrome, bronchial and pulmonary edema, bronchitis orpneumonitis, eosinophilic cellulites (e.g., Well's syndrome),eosinophilic pneumonias (e.g., Loeffler's syndrome, chronic eosinophilicpneumonia), eosinophilic fasciitis (e.g., Shulman's syndrome),delayed-type hypersensitivity, non-allergic asthma; exercise inducedbronchoconstriction; chronic obstructive pulmonary disease (COPD), acutebronchitis, chronic bronchitis, cough, pulmonary emphysema; systemicanaphylaxis or hypersensitivity responses, drug allergies (e.g., topenicillin, cephalosporin), eosinophiliamyalgia syndrome due to theingestion of contaminated tryptophane, insect sting allergies;autoimmune diseases, such as rheumatoid arthritis, Graves' disease,Sjogren's syndrome psoriatic arthritis, multiple sclerosis, systemiclupus erythematosus, myasthenia gravis, immune thrombocytopenia (adultITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic anemia(auto-immune and drug induced), Evans syndrome (platelet and red cellimmune cytopaenias), Rh disease of the newborn, Goodpasture's syndrome(anti-GBM disease), Celiac, autoimmune cardio-myopathy juvenile onsetdiabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease;graft rejection (e.g., in transplantation), including allograftrejection or graft versus-host disease; inflammatory bowel diseases,such as Crohn's disease and ulcerative colitis; spondyloarthropathies;scleroderma; psoriasis (including T-cell mediated psoriasis) andinflammatory dermatoses such as an dermatitis, eczema, atopicdermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g.,necrotizing, cutaneous, and hypersensitivity vasculitis); erythemanodosum; eosinophilic myositis, eosinophilic fasciitis, cancers withleukocyte infiltration of the skin or organs; ophthalmic diseases suchas age related macular degeneration, diabetic retinopathy and diabeticmacular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis,vernal keratoconjunctivitis, scarring, anterior segment scarring,blepharitis, blepharoconjunctivitis, bullous disorders, cicatricialpemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye,episcleritis, glaucoma, gliosis, Granuloma annulare, Graves'ophthalmopathy, intraocular melanoma, Pinguecula, proliferativevitreoretinopathy, pterygia, scleritis, uveitis, acute gout flares, goutor osteoarthritis.

(5) Pain such as chronic idiopathic pain syndrome, neuropathic pain,dysesthesia, allodynia, migraine, dental pain and post-surgical pain.

(6) Depression, anxiousness, diabetic neuropathy and bladder disorderssuch as bladder outlet obstruction, overactive bladder, cystitis;myocardial reperfusion injury or brain ischaemia injury.

In a further aspect the present invention relates to a compound ofgeneral formula 1 for use in the treatment and/or prevention of abovementioned diseases and conditions.

In a further aspect the present invention relates to the use of acompound of general formula 1 for the preparation of a medicament forthe treatment and/or prevention of above mentioned diseases andconditions.

In a further aspect of the present invention the present inventionrelates to methods for the treatment or prevention of above mentioneddiseases and conditions, which method comprises the administration of aneffective amount of a compound of general formula 1 to a human being.

Combination Therapy

The compounds of the invention may further be combined with one or more,preferably one additional therapeutic agent. According to one embodimentthe additional therapeutic agent is selected from the group oftherapeutic agents useful in the treatment of diseases or conditionsdescribed hereinbefore, in particular associated with fibrotic diseases,inflammatory and immunoregulatory disorders, respiratory orgastrointestinal diseases or complaints, inflammatory diseases of thejoints or of the nasopharynx, eyes, and skin or conditions such as forexample cough, idiopathic pulmonary fibrosis, other pulmonaryinterstitial diseases, asthma or allergic diseases, eosinophilicdiseases, chronic obstructive pulmonary disease, atopic dermatitis aswell as autoimmune pathologies, such as rheumatoid arthritis andatherosclerosis, or therapeutic agents useful for the treatment ofophthalmic diseases, pain and depression.

Additional therapeutic agents that are suitable for such combinationsinclude in particular those, which, for example, potentiate thetherapeutic effect of one or more active substances with respect to oneof the indications mentioned and/or allow the dosage of one or moreactive substances to be reduced.

Therefore, a compound of the invention may be combined with one or moreadditional therapeutic agents selected from the group consisting ofantifibrotic agents, anti-tussive agents, anti-inflammatory agents,anti-atopic dermatitis agents, analgesics, anti-convulsants,anxiolytics, sedatives, skeletal muscle relaxants or anti-depressants.

Antifibrotic agents are for example nintedanib, pirfenidone,phosphodiesterase-IV (PDE4) inhibitors such as roflumilast, autotaxininhibitors such as GLPG-1690 or BBT-877; connective tissue growth factor(CTGF) blocking antibodies such as Pamrevlumab; B-cell activating factorreceptor (BAFF-R) blocking antibodies such as Lanalumab; alpha-V/beta-6blocking inhibitors such as BG-00011/STX-100, recombinant pentraxin-2(PTX-2) such as PRM-151; c-Jun N-terminal kinase (JNK) inhibitors suchas CC-90001; galectin-3 inhibitors such as TD-139; G-protein coupledreceptor 84 (GPR84) inhibitors such as GLPG-1205; G-protein coupledreceptor 84/G-protein coupled receptor 40 dual inhibitors such asPBI-4050; Rho Associated Coiled-Coil Containing Protein Kinase 2 (ROCK2)inhibitors such as KD-025; heat shock protein 47 (HSP47) smallinterfering RNA such as BMS986263/ND-L02-s0201; Wnt pathway inhibitorsuch as SM-04646; LD4/PDE3/4 inhibitors such as Tipelukast; recombinantimmuno-modulatory domains of histidyl tRNA synthetase (HARS) such asATYR-1923; prostaglandin synthase inhibitors such as ZL-2102/SAR-191801;15-hydroxy-eicosapentaenoic acid (15-HEPE e.g. DS-102); Lysyl OxidaseLike 2 (LOXL2) inhibitors such as PAT-1251, PXS-5382/PXS-5338;phosphoinositide 3-kinases (PI3K)/mammalian target of rapamycin (mTOR)dual inhibitors such as HEC-68498; calpain inhibitors such as BLD-2660;mitogen-activated protein kinase kinase kinase (MAP3K19) inhibitors suchas MG-S-2525; chitinase inhibitors such as OATD-01; mitogen-activatedprotein kinase-activated protein kinase 2 (MAPKAPK2) inhibitors such asMMI-0100; transforming growth factor beta 1 (TGF-beta1) smallinterfering RNA such as TRK250/BNC-1021; or lysophosphatidic acidreceptor antagonists such as BMS-986278.

Anti-tussive agents are, for example, purinoceptor 3 (P2X3) receptorantagonists such as gefapixant, S-600918, BAY-1817080, or BLU-5937;neurokinin 1 (NK-1) receptor antagonist such as Orvepitant, Aprepitant;nicotinic acetylcholine receptor alpha 7 subunit stimulator such asATA-101/bradanicline; codeine, gabapentin, pregablin, or azithromycin.Anti-inflammatory agents are, for example, corticosteroids such asprednisolone or dexamethasone; cyclo-oxygenase-2 (COX2) inhibitors suchas celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib orlumiracoxib; prostaglandin E2 antagonists; leukotriene B4 antagonists;leukotriene D4 antagonists such as monteleukast; 5-lipoxygenaseinhibitors; or other nonsteroidal anti-inflammatory agents (NSAIDs) suchas aspirin, diclofenac, diflunisal, etodolac, ibuprofen or indomethacin.

Anti-atopic dermatitis agents are, for example, cyclosporin,methotrexate, mycophenolate mofetil, azathioprine, phosphodiesteraseinhibitors (e.g. apremilast, crisaborole), Janus Associated Kinase (JAK)inhibitors (e.g. tofacitinib), neutralizing antibodies againstIL-4/IL-13 (e.g. dupilamab), IL-13 (e.g. lebrikizumab, tralokinumab) andIL-31 (nemolizumab). Analgesics are, for example, of the opioid type,such as morphine, oxymorphine, levopanol, oxycodon, propoxyphene,nalmefene, fentanyl, hydrocondon, hydromorphone, meripidine, methadone,nalorphine, naloxone, naltrexone, buprenorphine, butorphanol,nalbuphine, pentazocine; or of the non-opioid type, such asacetophenamine Anti-depressants are, for example, tricyclicanti-depressants such as amitriptyline, clomipramine, despramine,doxepin, desipramine, imipramine, nortriptyline; selective serotoninreuptake inhibitor anti-depressants (SSRIs) such as fluoxetine,paroxetine, sertraline, citalopram, escitalopram; norepinephrinereuptake inhibitor anti-depressants (SNRIs) such as maprotiline,lofepramine, mirtazapine, oxaprotiline, fezolamine, tomoxetine,mianserin, buproprion, hydroxybuproprion, nomifensine, viloxazine; dualserotonin-norepinephrine reuptake inhibitor anti-depressants (SNRIs)such as duloxetine, venlafaxine, desvenlafaxine, levomilnacipran;atypical antidepressants such as trazodone, mirtazapine, vortioxetine,vilazodone, bupropion; or monoamine oxidase inhibitor anti-depressantss(MAOIs) such as tranylcypromine, phenelzine, or isocarboxazid.

Anxiolytics are, for example, benzodiazepines such as alprazolam,bromazepam, chlordiazepoxide, clonazepam, clorazepate, diazepam,flurazepam, lorazepam, oxazepam, temazepam, triazolam, or tofisopam; orthey are nonbenzodiazepine hypnoticssuch as eszopiclone, zaleplon,zolpidem, or zopiclone; or they are carbamates e.g. meprobamate,carisoprodol, tybamate, or lorbamate; or they are antihistamines such ashydroxyzine, chlorpheniramine or diphenhydramine.

Sedatives are, for example, barbiturate sedatives, such as amobarbital,aprobarbital, butabarbital, butabital, mephobarbital, metharbital,methohexital, pentobarbital, secobarbital, talbutal, theamylal, orthiopental; or they are non-barbiturate sedatives such as glutethimide,meprobamate, methaqualone or dichloalphenazone.

Skeletal muscle relaxants are, for example, baclofen, meprobamate,carisoprodol, cyclobenzaprine, metaxalone, methocarbamol, tizanidine,chlorzoxazone or orphenadrine.

Other suitable combination partners are inhibitors ofAcetylcholinesterase inhibitors such as donepezil; 5-HT-3 anatgonistssuch as ondansetron; metabotropic glutamate receptor antagonists;antiarrhythmics such as mexiletine or phenytoin; or NMDA receptorantagonists. Further suitable combination partners are incontinencemedications, for example, antichos linergics such as oxybutynin,tolterodine, darifenacin, fesoterodine, solifenacin or trospium; or theyare bladder muscle relaxants such as mirabegron; or they are alphablockers such as tamsulosin, alfuzosin, silodosin, doxazosin orterazosin.

The dosage for the combination partners mentioned above is usually ⅕ ofthe lowest dose normally recommended up to 1/1 of the normallyrecommended dose.

Therefore, in another aspect, this invention relates to the use of acompound according to the invention in combination with one or moreadditional therapeutic agents described hereinbefore and hereinafter forthe treatment of diseases or conditions which may be affected or whichare mediated by TRPA1, in particular diseases or conditions as describedhereinbefore and hereinafter.

In a further aspect this invention relates to a method for treating adisease or condition which can be influenced by the inhibition of TRPA1in a patient that includes the step of administering to the patient inneed of such treatment a therapeutically effective amount of a compoundof formula (I) or a pharmaceutically acceptable salt thereof incombination with a therapeutically effective amount of one or moreadditional therapeutic agents.

In a further aspect this invention relates to the use of a compound offormula (I) or a pharmaceutically acceptable salt thereof in combinationwith one or more additional therapeutic agents for the treatment ofdiseases or conditions which can be influenced by the inhibition ofTRPA1 in a patient in need thereof.

In yet another aspect the present invention relates a method for thetreatment of a disease or condition mediated by TRPA1 activity in apatient that includes the step of administering to the patient,preferably a human, in need of such treatment a therapeuticallyeffective amount of a compound of the present invention in combinationwith a therapeutically effective amount of one or more additionaltherapeutic agents described in hereinbefore and hereinafter.

The use of the compound according to the invention in combination withthe additional therapeutic agent may take place simultaneously or atstaggered times.

The compound according to the invention and the one or more additionaltherapeutic agents may both be present together in one formulation, forexample a tablet or capsule, or separately in two identical or differentformulations, for example as a so-called kit-of-parts. Consequently, inanother aspect, this invention relates to a pharmaceutical compositionthat comprises a compound according to the invention and one or moreadditional therapeutic agents described hereinbefore and hereinafter,optionally together with one or more inert carriers and/or diluents.

In yet another aspect the present invention relates to the use of acompound according to the invention in a cough-measuring device.

Other features and advantages of the present invention will becomeapparent from the following more detailed examples which illustrate, byway of example, the principles of the invention.

Preparation

The compounds according to the present invention and their intermediatesmay be obtained using methods of synthesis which are known to the oneskilled in the art and described in the literature of organic synthesis.Preferably, the compounds are obtained in analogous fashion to themethods of preparation explained more fully hereinafter, in particularas described in the experimental section. In some cases, the order incarrying out the reaction steps may be varied. Variants of the reactionmethods that are known to the one skilled in the art but not describedin detail here may also be used.

The general processes for preparing the compounds according to theinvention will become apparent to the one skilled in the art studyingthe following schemes. Starting materials may be prepared by methodsthat are described in the literature or herein, or may be prepared in ananalogous or similar manner Any functional groups in the startingmaterials or intermediates may be protected using conventionalprotecting groups. These protecting groups may be cleaved again at asuitable stage within the reaction sequence using methods familiar tothe one skilled in the art.

The compounds according to the invention are prepared by the methods ofsynthesis described hereinafter in which the substituents of the generalformulae have the meanings given hereinbefore. These methods areintended as an illustration of the invention without restricting itssubject matter and the scope of the compounds claimed to these examples.Where the preparation of starting compounds is not described, they arecommercially obtainable or may be prepared analogously to knowncompounds or methods described herein. Substances described in theliterature are prepared according to the published methods of synthesis.

Compounds of formula I may be prepared as shown in Scheme I below.

In scheme I, chloromethyltetrazole is N-alkylated with an appropriatearomatic or heteroaromatic acetophenone derivative carrying a leavinggroup “LG” (e.g. Cl or Br) alpha to the carbonyl group. The reaction cantypically be run at ambient temperature or 50° C. in the presence of abase (e.g. K₂CO₃). The carbonyl group of side chain (A) can be reducedin an enantioselective fashion by using appropriate catalytic systemsusing a transition metal complex (of e.g. Ru or IR) in combination witha chiral ligand (e.g.([(1S,2S)-(−)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido). Inthe presence of a base (e.g. K₂CO₃) side chain (B) can be used asalkylating agent for various thienopyrimidinones to afford the compoundsof general formula (I).

Tricyclic thienopyrimidone compounds may be prepared as shown in SchemeII below.

In scheme II, a suitable thieno-tetrahydropyridine precursor isconverted with an appropriate reagent such as formamidine, formamide ora salt thereof in a suitable solvent (e.g. EtOH) at elevatedtemperatures (e.g. 100° C.) into the tricyclic thieno-pyrimidonecompound (C). The tetrahydropyridine core is then oxidized using asuitable oxidant (e.g. KMnO₄) and is accelerated by the presence of achelating reagent (e.g. 18-crown-6). This reaction is typicallyperformed in a non-polar solvent (e.g. DCM) and run preferentially atambient temperature. In case R³ resembles a protecting group (e.g. BOC),this group can be removed using suitable conditions for deprotection(e.g. TFA/DCM or HCl/dioxane at RT for R³=BOC).

EXAMPLES

Preparation

The compounds according to the invention and their intermediates may beobtained using methods of synthesis which are known to the one skilledin the art and described in the literature of organic synthesis forexample using methods described in “Comprehensive OrganicTransformations”, 2nd Edition, Richard C. Larock, John Wiley & Sons,2010, and “March's Advanced Organic Chemistry”, 7th Edition, Michael B.Smith, John Wiley & Sons, 2013. Preferably the compounds are obtainedanalogously to the methods of preparation explained more fullyhereinafter, in particular as described in the experimental section. Insome cases the sequence adopted in carrying out the reaction schemes maybe varied. Variants of these reactions that are known to the skilledartisan but are not described in des tail herein may also be used. Thegeneral processes for preparing the compounds according to the inventionwill become apparent to the skilled man on studying the schemes thatfollow. Starting compounds are commercially available or may be preparedby methods that are described in the literature or herein, or may beprepared in an analogous or similar manner Before the reaction iscarried out, any corresponding functional groups in the startingcompounds may be protected using conventional protecting groups. Theseprotecting groups may be cleaved again at a suitable stage within thereaction sequence using methods familiar to the skilled man anddescribed in the literature for example in “Protecting Groups”, 3rdEdition, Philip J. Kocienski, Thieme, 2005, and “Protective Groups inOrganic Synthesis”, 4th Edition, Peter G. M. Wuts, Theodora W. Greene,John Wiley & Sons, 2006. The terms “ambient temperature” and “roomtemperature” are used interchangeably and designate a temperature ofabout 20° C., e.g. between 19 and 24° C.

Abbreviations:

ACN acetonitrile Aq. aqueous ° C. Degree celsius CyH/CH cyclohexaneconc. concentrated DCM dichloro methane DIPEA N,N-diisopropylethylamineDMA N,N-dimethylacetamide DMF N,N-dimethylformamide DMSO dimethylsulfoxide ESI-MS Electrospray ionisation mass spectrometry EtOAc ethylacetate EtOH ethanol ex example eq equivalent h hour HCl Hydrochloridacid HPLC High performance liquid chromatography K₂CO₃ potassiumcarbonate L liter M molar MeOH methanol MgSO₄ magnesium sulphate minminute mL milliliter MTBE tert-butylmethylether n.d. not determined NH₃ammonia RT room temperature (about 20° C.) sat. saturated TBTUBenzotriazolyl tetramethyluronium tetrafluoroborate TEA triethylamineTFA trifluoroacetic acid THF tetrahydrofuran

Preparation of Starting Compounds

Intermediate I Intermediate I.1 (General Route)1-(4-bromophenyl)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]ethan-1-one

To 1.00 g (8.44 mmol) of 5-(chloromethyl)-2H-1,2,3,4-tetrazole and 2.17g (9.28 mmol) 4-bromide in 15 mL DMA are added 1.63 g (11.8 mmol) K₂CO₃.The reaction mixture is stirred at RT for 30 min and before the mixtureis filtered. The solution is diluted with water and sat. aq.NaCl-solution and is extracted three times with EtOAc. The combinedorganic phase is washed with water, dried over Na₂SO₄, filtered overactivated charcoal and the solvent is removed in vacuo. The residue ispurified by column chromatography (silica gel; CH/EtOAc, 8/2→1/1).

C₁₀H₈Cl₂N₄O (M=271.1 g/mol)

ESI-MS: 271 [M+H]⁺

R_(t) (HPLC): 1.01 min (method B)

The following compounds are prepared according to the general procedure(intermediate I.1) described above:

HPLC reten- tion time (method) Int. Starting materials Structure ESI-MS[min] I.2

315/317 [M + H]⁺ 1.00 (C) I.3

311 [M + H]⁺ 1.08 (C) I.4

277 [M + H]⁺ 1.01 (C) I.5

273 [M + H]⁺ 1.00 (C) I.6 VII.1

311 [M + H]⁺ 1.02 (B) I.7

251 [M + H]⁺ 0.95 (C) I.8

281 [M + H]⁺ 0.89 (C) I.9 VII.2

295 [M + H]⁺ 1.02 (B)  I.10 VII.3

295 [M + H]⁺ 1.02 (B)  I.11 VII.4

268 [M + H]⁺ 0.47 (G)  I.12

293 [M + H]⁺ 1.12 (B)

Intermediate II Intermediate II.1 (General Route)(1R)-1-(4-chlorophenyl)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]ethan-1-ol

1.30 g (4.80 mmol)1-(4-chlorophenyl)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]ethan-1-one(example I.1) are added to 20 mL ACN under inert atmosphere. 11.9 mg(0.02 mmol)Chloro([1S,2S)-(−)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido)(mesitylene)ruthenium(II) (CAS 174813-81-1) are added before 0.72 mL (1.73 mmol) formic acidtriethylamine complex (5:2) are added dropwise. After stirring at RT for3 h the solvent is removed in vacuo. To the remaining crude mixture isadded water and this mixture is extracted with EtOAc. The organic layersare combined, dried over Na₂SO₄ and activated charcoal, filtered and thesolvent is removed in vacuo.

C₁₀H₁₀Cl₂N₄O (M=273.1 g/mol)

ESI-MS: 273 [M+H]⁺

R_(t) (HPLC): 0.96 min (method B)

The following compounds are prepared according to the general procedure(intermediate II.1) described above:

HPLC reten- tion time (method) Int. Starting materials Structure ESI-MS[min] II.2 I.2

317/319 [M + H]⁺ 1.14 (B) II.3 I.3

313 [M + H]⁺ 1.03 (B) II.4 I.4

279 [M + H]⁺ 0.97 (C) II.5 I.5

275 [M + H]⁺ 0.48 (A) II.6 I.6

313 [M + H]⁺ 1.01 (B) II.7 I.7

253 [M + H]⁺ 0.48 (A) II.8 I.8

283 [M + H]⁺ 0.43 (A) II.9 I.9

297 [M + H]⁺ 0.97 (B)  II.10  I.10

297 [M + H]⁺ 0.97 (B)  II.11  I.11

270 [M + H]⁺ 0.41 (A)  II.12  I.12

295 [M + H]⁺ 1.10 (B)

Intermediate III Intermediate III.1 (General Route) Tert-butyl4,8-dioxo-5,6-dihydro-3H-pyrido[2,3]thieno[2,4-c]pyrimidine-7-carboxylate

500 mg (1.63 mmol) tert-butyl 4-oxo-3,5,6,8-tetrahydropyrido [2,3]thieno[2,4-c]pyrimidine-7-carboxylate, 514 mg (3.25 mmol) potassiumpermanganate, 86.0 mg (0.33 mmol) 18-crown-6 are added to 8 mL DCM. Thereaction mixture is stirred at RT overnight. The mixture is diluted withMeOH and with 10% sodium metabisulfite solution. The precipitate isfiltered off and the solution is evaporated. The crude residue issuspended in MeOH/DMF, filtered and purified by HPLC (ACN/H₂O/TFAgradient).

C₁₄H₁₅N₃O₄S (M=321.4 g/mol)

ESI-MS: 322 [M+H]⁺

R_(t) (HPLC): 0.45 min (method A)

The following compounds are prepared according to the general procedure(example III.1) described above:

HPLC reten- tion time Starting ESI- (method) Ex. materials Structure MS[min] III.2 V.1

236 [M + H]⁺ 0.27 (A)

Intermediate IV Intermediate IV.1 (General Route)6,7-dihydropyrido[4′,3′:4,5]thieno[2,3-d]pyrimidine-4,8(3H,5H)-dione

14.0 mg (0.04 mmol) tert-butyl4,8-dioxo-5,6-dihydro-3H-pyrido[2,3]thieno[2,4-c]pyrimidine-7-carboxylate(example III.1) in 500 μL DCM is treated with 35 μL (0.45 mmol) TFA. Thereaction mixture is stirred at RT for 0.5 h and the mixture isconcentrated to give the desired product that was used in the next stepwithout further purification.

C₉H₇N₃O₂S (M=221.2 g/mol)

ESI-MS: 222 [M+H]⁺

R_(t) (HPLC): 0.22 min (method A)

Intermediate V Intermediate V.1 (General Route)7-methyl-3,5,6,8-tetrahydropyrido[2,3]thieno[2,4-c]pyrimidin-4-one

1.00 g (4.00 mmol) ethyl 2-amino-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylate in 10 mL EtOH is treated with 2.89 mL(31.0 mmol) formamidine acetate. The reaction mixture is stirred at 100°C. for 6 h. After that, 0.58 mL (19.0 mmol) formamidine acetate is addedand the mixture is stirred another 4h at 100° C. After cooling toambient temperature, the product is filtered off and is washed withethanol. The collected precipitate is dried in an oven at 50° C.

C₁₀H₁₁N₃OS (M=221.3 g/mol)

ESI-MS: 222 [M+H]⁺

R_(t) (HPLC): 0.15 min (method A)

Intermediate VI Intermediate VI.1 (General Route)1-(6-chloro-1-benzofuran-2-yl)ethan-1-one

1.00 g (6.39 mmol) 4-chloro-2-hydroxybenzaldehyde in 10 mL acetone underargon is treated with 0.56 mL (7.03 mmol) chloroacetone in 10 mL acetoneis treated with 1.47 g (10.6 mmol) potassium carbonate and 2.89 mL (31.0mmol) formamidine acetate under argon. The reaction mixture is stirred3h at 70° C. The reaction mixture is cooled down, the precipitate isfiltered off and the solution is evaporated. The residue isrecrystallized in 10 mL ice cold MeOH, the crystals are filtered off anddried overnight.

C₁₀H₇ClO₂ (M=194.6 g/mol)

EI-MS: 194 [M*]⁺

R_(t) (HPLC): 1.02 min (method B)

The following compounds are prepared according to the general procedure(intermediate VI.1) described above:

HPLC reten- tion time (method) Int. Starting materials Structure ESI-MS[min] VI..2

179 [M + H]⁺ 0.95 (B) VI..3

179 [M + H]⁺ 0.95 (B)

Intermediate VII Intermediate VII.1 (General Route)2-bromo-1-(6-chloro-1-benzofuran-2-yl)ethan-1-one

840 mg (4.32 mmol) 1-(6-chloro-1-benzofuran-2-yl)ethan-1-one (exampleVI.1) in 10 mL THF is treated with 2.08 g (4.32 mmol) tetrabutylammoniumtribromide, dissolved in 500 μL MeOH and 5 mL THF, dropwise. Thereaction mixture is stirred for 1 h at RT. The reaction mixture isevaporated, the residue is diluted with water/1M HCl and is extractedwith MTBE. The organic phases are combined, washed with 1M HCl andwater, dried over MgSO4, filtered and the solvent is removed in vacuo.The residue is purified by column chromatography (silica gel; CH/EtOAc,95/5 to 50/50).

C₁₀H₆BrClO₂ (M=273.5 g/mol)

ESI-MS: 273/275 [M+H]⁺

R_(t) (HPLC): 1.10 min (method B)

The following compounds are prepared according to the general procedure(intermediate VII.1) described above:

HPLC reten- tion time (method) Int. Starting materials Structure ESI-MS[min] VII..2 VI.2

257/259 [M + H]⁺ 1.03 (B) VII..3 VI.3

Mass not de- tected 1.03 (B) VII.4*

228/230 [M − H]⁻ 0.75 (H) *The reaction is performed with bromine (13.6eq) at RT for 2 h in dioxane/diethyl ether and quenched with sodiumthiosulfate solution.

Intermediate VIII Intermediate VIII.1 (General Route)5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxylic Acid

2.00 g (8.92 mmol) methyl5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxylate in 25 mL MeOHis treated with 6.69 mL (26.8 mmol) 4M aq. NaOH solution. The reactionmixture is stirred at RT for 2h. The solution is then acidified byadding 4M aq. HCl solution, the crystals are filtered, washed with waterand dried in the oven at 60° C. to give the desired product.

C₈H₆N₂O₃S (M=210.2 g/mol)

ESI-MS: 211 [M+H]⁺

R_(t) (HPLC): 0.29 min (method E)

Intermediate IX Intermediate IX.1 (General Route)N,5-dimethyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxamide

250 mg (1.19 mmol)5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxylic acid (exampleVIII.1) in 30 mL THF is treated with 429 mg (1.19 mmol) TBTU and 331 μL(2.38 mmol) TEA. The reaction mixture is stirred at RT for 30 min before654 μL (1.31 mmol) methylamine (2M solution in THF) is added to thesolution. The reaction mixture is stirred at RT overnight. The reactionmixture is evaporated, the residue is extracted with DCM/water, theorganic phases are combined, dried, filtered and the solvent is removedin vacuo. The residue is purified by column chromatography (silica gel;DCM/MeOH/NH₃ 90/10/1).

C₉H₉N₃O₂S (M=223.3 g/mol)

ESI-MS: 224 [M+H]⁺

R_(t) (HPLC): 0.62 min (method C)

Intermediate X Intermediate X.1 (General Route)

To 5.00 g (26.8 mmol) 2-amino-4-chloro-3-cyano-5-formylthiophene (CAS:104366-23-6) in 25 mL pyridine is added 5.70 mL (42.9 mmol)N,N-dimethylformamide dimethyl acetal. The mixture is stirred at 100° C.for 3 h. After cooling to ambient temperature, the mixture isconcentrated under reduced pressure. The residue is resuspended in DCMand washed with water. The organic phase is dried over Na₂SO₄ andconcentrated to yield the desired product.

C₉H₈ClN₃OS (M=241.7 g/mol)

ESI-MS: 242 [M+H]⁺

R_(t) (HPLC): 1.04 min (method B)

Intermediate XI Intermediate XI.1 (General Route)

To 1.00 g (4.14 mmol) of example X.1 in 10 mL formic acid is added 678mg (8.27 mmol) sodium acetate. The reaction mixture is stirred at refluxovernight. After cooling to ambient temperature, the mixture is pouredonto ice-cold water. It is then diluted with dichloromethane andconcentrated to dryness. The residue is suspended in dichloromethane andthe remaining salts are filtered off. The filtrate is concentrated toyield the desired product. C₇H₃ClN₂O₂S (M=214.6 g/mol)

ESI-MS: 213 [M−H]⁻

R_(t) (HPLC): 0.87 min (method B)

Intermediate XII Intermediate XII.1 (General Route)

To 150 mg (699 μmol) of example XI.1 in 4.0 mL DMF is added 473 mg (769μmol) potassium peroxymonosulfate. The reaction mixture is added atambient temperature for 18 h. The mixture is then purified bypreparative HPLC (H₂O/ACN/TFA) to yield the desired product.

C₇H₃ClN₂O₃S (M=230.6 g/mol)

ESI-MS: 229 [M−H]⁻

R_(t) (HPLC): 0.58 min (method C)

Intermediate XIII Intermediate XIII 1 (General Route)

To 45.0 mg (0.195 mmol) of example XII.1 in 1.0 mL DMF is added 81.7 mg(0.215 mmol)1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate, 74.8 μL (0.429 mmol) diisopropylethylamineand 1.17 mL (0.5 M, 0.585 mmol) of a solution of ammonia in THF. Thereaction mixture is stirred at ambient temperature for 18 h. Thereaction mixture is then purified by preparative HPLC (H₂O/ACN/NH₃gradient) to yield the desired product.

C₇H₄ClN₃O₂S (M=229.6 g/mol)

ESI-MS: 230 [M+H]⁺

R_(t) (HPLC): 0.52 min (method C)

Preparation of Final Compounds

Example 1 (General Route)3-({2-[(2R)-2-(4-chlorophenyl)-2-hydroxyethyl]-2H-1,2,3,4-tetrazol-5-yl}methyl)-5-methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxamide

919 mg (4.39 mmol) of5-Methyl-4-oxo-3H,4H-thieno[2,3-d]pyrimidine-6-carboxamide are added to40 mL DMA. Then 1.97 g (14.3 mmol) K₂CO₃ and 1.20 g (4.39 mmol) ofexample II.1 are added and the mixture is stirred at 50° C. for 3 h.After cooling to RT, the mixture is purified by HPLC (ACN/H₂O/TFAgradient) to yield the desired product.

C₁₈H₁₆ClN₇O₃S (M=445.9 g/mol)

ESI-MS: 446 [M+H]⁺

R_(t) (HPLC): 0.92 min (method B)

¹H NMR (400 MHz, DMSO-d₆) δ ppm: 2.70 (s, 3H), 4.73-4.82 (m, 2H), 5.12(dt, J=7.4, 5.1 Hz, 1H), 5.49 (s, 2H), 5.91 (d, J=4.8 Hz, 1H), 7.33-7.41(m, 4H), 7.67 (br s, 2H), 8.66 (s, 1H).

The following compounds are prepared according to the general procedure(example 1.1) described above:

Starting mate- Reaction Ex. rials Structure conditions 2 II.2 +5-Methyl-4- oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

Solvent: DMA, RT 20 h 3 II.3 + 5-Methyl-4- oxo-3H,4H- thieno[2,3-d]pyrimidine- 6-carboxamide

Solvent: DMF; 4 II.4 + 5-Methyl-4- oxo-3H,4H- thieno[2,3- d]pyrimidine-6-carboxamide

Solvent: DMF; di- rectly puri- fied by HPLC 5 II.5 + 5-Methyl-4-oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

Solvent: DMF; di- rectly puri- fied by HPLC 6 II.2 + IV.1

3.2eq II.2, 6.1eq base; 3 h RT 7 II.2 + III.2

Solvent: DMF; 3 h RT; 2 h 50° C. 8 II.6 + 5-Methyl-4- oxo-3H,4H-thieno[2,3- d]pyrimidine- 6-carboxamide

Solvent: DMF; 3 h 50° C.; di- rectly puri- fied by HPLC 9 II.7 +5-Methyl-4- oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

Solvent: DMF; over- night RT; directly pu- rified by HPLC 10 II.8 +5-Methyl-4- oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

Solvent: DMF; 2 h RT; directly purified by HPLC 11 II.9 + 5-Methyl-4-oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

3 h 50° C.; overnight RT; directly purified by HPLC 12 II.10 +5-Methyl-4- oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

3 h 50° C.; RT over- night; di- rectly puri- fied by HPLC 13 II.2 + iX.1

Solvent: DMF; 3 h RT; directly purified by HPLC 14 II.8 + IX.1

Solvent: DMF; 2 h RT; directly purified by HPLC 15 II.7 + IX.1

Solvent: DMF; 2 h RT; directly purified by HPLC 16 II.4 + IX.1

Solvent: DMF; 2 h RT; directly purified by HPLC 17 II.4 + IV.1

DMF, RT, 18 h; directly pu- rified by HPLC 18 II.3 + IV.1

DMF, RT, 18 h; directly pu- rified by HPLC 19 II.9 + IV.1

DMF, RT, 5.5 h; directly pu- rified by HPLC 20 II.7 + IV.1

DMF, RT, 19 h; directly pu- rified by HPLC 21 II.11 + IV.1

DMF, RT, 18 h; directly pu- rified by HPLC 22 II.12 + 5-Methyl-4-oxo-3H,4H- thieno[2,3- d]pyrimidine- 6-carboxamide

DMF, 3 h 50° C., over- night RT; directly pu- rified by HPLC 23 II.2 +XIII.1

DMF, 18 h RT; directly pu- rified by HPLC

Analytical data for the compounds described in the table above:

HPLC retention time Ex. ESI-MS (method) [min] ¹H NMR (400 MHz, DMSO-d₆)δ ppm 2 490/492 [M + H]⁺ 1.02 (B) 2.70 (s, 3H), 4.73-4.83 (m, 2H), 5.10(dd, J = 7.5, 5.3 Hz, 1H), 5.49 (s, 2H), 5.95 (br s, 1H), 7.29-7.35 (m,2H), 7.47-7.52 (m, 2H), 7.67 (s, 2H), 8.66 (s, 1H). 3 486 [M + H]⁺ 0.63(D) 2.69 (s, 3H), 4.97-5.08 (m, 2H), 5.27 (dt, J = 7.6, 5.5 Hz, 1H),5.48 (s, 2H), 6.29 (d, J = 5.8 Hz, 1H), 6.82 (s, 1H), 7.30 (dd, J = 8.7,2.3 Hz, 1H), 7.58 (d, J = 8.6 Hz, 1H), 7.63-7.69 (m, 3H), 8.63 (s, 1H) 4452 [M + H]⁺ 0.87 (C) 2.69 (s, 3H), 4.81 (dd, J = 13.6, 8.1 Hz, 1H),4.89 (dd, J = 13.7, 4.6 Hz, 1H), 5.25-5.32 (m, 1H), 5.50 (s, 2H), 6.39(d, J = 5.3 Hz, 1H), 6.86 (dd, J = 3.8, 0.8 Hz, 1H), 6.93 (d, J = 3.8Hz, 1H), 7.66 (s, 2H), 8.66 (s, 1H) 5 448 [M + H]⁺ 0.84 (C) 2.69 (s,3H), 4.75-4.85 (m, 2H), 5.13 (dd, J = 7.4, 5.3 Hz, 1H), 5.49 (s, 2H),5.98 (br s, 1H), 7.18-7.24 (m, 1H), 7.35 (dt, J = 10.7, 8.4 Hz, 1H),7.45 (ddd, J = 11.7, 7.9, 2.0 Hz, 1H), 7.66 (s, 2H), 8.65 (s, 1H) 6 458[M + H]⁺ 0.44 (A) δ ppm 3.12 (t, J = 7.0 Hz, 2H), 3.50 (td, J = 7.1, 2.5Hz, 2H), 4.73-4.84 (m, 2H), 5.12 (dd, J = 7.4, 5.3 Hz, 1H), 5.51 (s,2H), 6.28 (br s, 1H), 7.34-7.41 (m, 4H), 7.95 (br s, 1H), 8.70 (s, 1H) 7472 [M + H]⁺ 0.49 (A) 2.99 (s, 3H), 3.20 (t, J = 7.2 Hz, 2H), 3.66 (t, J= 7.1 Hz, 2H), 4.72-4.83 (m, 2H), 5.12 (dd, J = 7.4, 5.5 Hz, 1H), 5.50(s, 2H), 5.67-6.28 (br s, 1H), 7.31-7.45 (m, 4H), 8.69 (s, 1H) 8 486[M + H]⁺ 0.63 (D) 2.68 (s, 3H), 4.99 (dd, J = 13.8, 7.5 Hz, 1H), 5.04(dd, J = 13.8, 5.1 Hz, 1H), 5.26 (dt, J = 7.3, 5.4 Hz, 1H), 5.48 (s,2H), 6.29 (d, J = 5.7 Hz, 1H), 6.85 (s, 1H), 7.27 (dd, J = 8.3, 1.8 Hz,1H), 7.58 (d, J = 8.4 Hz, 1H), 7.66 (br s, 2H), 7.69-7.74 (m, 1H), 8.62(s, 1H) 9 426 [M + H]⁺ 0.41 (A) 2.26 (s, 3H), 2.70 (s, 3H), 4.73 (d, J =6.5 Hz, 2H), 5.05 (t, J = 6.5 Hz, 1H), 5.3-6.1 (br s, 1H), 5.49 (s, 1H),7.08-7.13 (m, J = 7.9 Hz, 2H), 7.21-7.26 (m, J = 8.0 Hz, 2H), 7.66 (s,2H), 8.65 (s, 1H) 10 456 [M + H]⁺ 0.80 (C) 2.69 (s, 3H), 4.71 (dd, J =13.5, 5.2 Hz, 2H), 4.75 (dd, J = 13.6, 7.6 Hz, 1H), 5.02 (dt, J = 7.3,5.1 Hz, 1H), 5.49 (s, 2H), 5.75 (d, J = 4.8 Hz, 1H), 5.96-5.99 (m, 2H),6.78-6.83 (m, 2H), 6.98 (s, 1H), 7.66 (s, 2H), 8.65 (s, 1H) 11 470 [M +H]⁺ 0.86 (B) 2.68 (s, 3H), 4.95-5.08 (m, 2H), 5.27 (dd, J = 7.7, 4.8 Hz,1H), 5.49 (s, 2H), 6.28 (br s, 1H), 6.83 (s, 1H), 7.11 (td, J = 9.2, 2.8Hz, 1H), 7.39 (dd, J = 8.9, 2.7 Hz, 1H), 7.56 (dd, J = 9.0, 4.2 Hz, 1H),7.68 (s, 2H), 8.63 (s, 1H) 12 470 [M + H]⁺ 0.56 (F) 2.68 (s, 3H),4.96-5.07 (m, 2H), 5.25 (dd, J = 7.6, 5.1 Hz, 1H), 5.49 (s, 2H),5.88-6.67 (br s, 1H), 6.83 (s, 1H), 7.10 (td, J = 9.3, 2.3 Hz, 1H), 7.50(dd, J = 9.3, 2.1 Hz, 1H), 7.58 (dd, J = 8.5, 5.6 Hz, 1H), 7.66 (s, 2H),8.63 (s, 1H) 13 460 [M + H]⁺ 0.61 (D) 2.67 (s, 3H), 2.77 (d, J = 4.6 Hz,3H), 4.73-4.83 (m, 2H), 5.11 (t, J = 6.3 Hz, 1H), 5.49 (s, 2H), 5.91 (s,1H), 7.33-7.42 (m, 4H), 8.16 (q, J = 4.1 Hz, 1H), 8.65 (s, 1H) 14 470[M + H]⁺ 0.84 (C) 2.67 (s, 3H), 2.77 (d, J = 4.6 Hz, 3H), 4.71 (dd, J =13.6, 5.4 Hz, 1H), 4.75 (dd, J = 13.7, 7.7 Hz, 1H), 5.02 (dd, J = 7.4,5.6 Hz, 1H), 5.49 (s, 2H), 5.75 (br s, 1H), 5.97-5.99 (m, 2H), 6.81 (d,J = 0.9 Hz, 2H), 6.97-6.99 (m, 1H), 8.16 (q, J = 4.2 Hz, 1H), 8.66 (s,1H) 15 440 [M + H]⁺ 0.89 (C) 2.26 (s, 3H), 2.67 (s, 3H), 2.77 (d, J =4.6 Hz, 3H), 4.70-4.78 (m, 2H), 5.05 (t, J = 6.5 Hz, 1H), 5.33-6.10 (brs, 1H), 5.49 (s, 2H), 7.11 (d, J = 7.9 Hz, 2H), 7.24 (d, J = 8.0 Hz,2H), 8.16 (q, J = 4.2 Hz, 1H), 8.65 (s, 1H) 16 466 [M + H]⁺ 0.90 (C)2.67 (s, 3H), 2.77 (d, J = 4.6 Hz, 3H), 4.81 (dd, J = 13.6, 8.1 Hz, 1H),4.89 (dd, J = 13.6, 4.6 Hz, 1H), 5.26-5.31 (m, 1H), 5.50 (s, 2H),5.92-6.73 (br s, 1H), 6.86 (dd, J = 3.8, 0.9 Hz, 1H), 6.93 (d, J = 3.8Hz, 1H), 8.16 (q, J = 4.3 Hz, 1H), 8.66 (s, 1H) 17 464 [M + H]⁺ 0.44 (A)3.12 (t, J = 7.0 Hz, 2H), 3.49 (td, J = 7.0, 2.5 Hz, 2H), 4.82 (dd, J =13.7, 8.2 Hz, 1H), 4.89 (dd, J = 13.7, 4.6 Hz, 1H), 5.29 (dd, J = 7.7,4.5 Hz, 1H), 5.52 (s, 2H), 6.39 (br s, 1H), 6.87 (dd, J = 3.8, 0.8 Hz,1H), 6.94 (d, J = 3.8 Hz, 1H), 7.92-7.97 (m, 1H), 8.70 (s, 1H) 18 498[M + H]⁺ 0.48 (A) 3.11 (t, J = 7.0 Hz, 2H), 3.50 (td, J = 7.1, 2.3 Hz,2H), 4.97-5.08 (m, 2H), 5.28 (dd, J = 7.7, 4.8 Hz, 1H), 5.50 (s, 2H),6.82 (s, 1H), 7.30 (dd, J = 8.7, 2.2 Hz, 1H), 7.58 (d, J = 8.7 Hz, 1H),7.66 (d, J = 2.3 Hz, 1H), 7.92-7.98 (m, 1H), 8.67 (s, 1H) 19 482 [M +H]⁺ 0.44 (A) 3.11 (t, J = 7.0 Hz, 2H), 3.50 (td, J = 7.1, 2.5 Hz, 2H),4.96-5.08 (m, 2H), 5.27 (dd, J = 7.7, 4.8 Hz, 1H), 5.50 (s, 2H), 6.28(d, J = 5.7 Hz, 1H), 6.83 (s, 1H), 7.11 (td, J = 9.2, 2.7 Hz, 1H), 7.39(dd, J = 8.9, 2.7 Hz, 1H), 7.56 (dd, J = 8.9, 4.1 Hz, 1H), 7.92-7.97 (m,1H), 8.67 (s, 1H) 20 438 [M + H]⁺ 0.43 (A) 2.26 (s, 3 H), 3.12 (t, J =7.0 Hz, 2H), 3.50 (td, J = 7.0, 2.5 Hz, 3H), 4.70-4.78 (m, 2H), 5.06 (t,J = 6.5 Hz, 1H), 5.52 (s, 2H), 5.54-5.99 (br s, 1H), 7.11 (d, J = 7.9Hz, 2H), 7.24 (d, J = 8.0 Hz, 2H), 7.91-8.00 (m, 1H), 8.70 (s, 1H) 21455 [M + H]⁺ 0.36 (A) 3.12 (t, J = 7.0 Hz, 2H), 3.50 (td, J = 7.1, 2.5Hz, 2H), 4.88 (dd, J = 13.7, 8.0 Hz, 1H), 4.99 (dd, J = 13.8, 4.3 Hz,1H), 5.46 (dd, J = 7.8, 3.9 Hz, 1H), 5.51 (s, 2H), 6.69 (br s, 1H), 7.18(dd, J = 3.9, 0.9 Hz, 1H), 7.81 (d, J = 3.9 Hz, 1H), 7.95 (s, 1H), 8.69(s, 1H) 22 468 [M + H]⁺ 0.60 (D) 2.70 (s, 3H), 4.91 (dd, J = 13.7, 8.4Hz, 1H), 4.98 (dd, J = 13.7, 4.7 Hz, 1H), 5.46 (dd, J = 7.9, 4.6 Hz,1H), 5.51 (s, 2 H), 6.43 (br s, 1H), 7.28-7.38 (m, 3H), 7.67 (s, 2H),7.71-7.76 (m, 1H), 7.89-7.94 (m, 1H), 8.66 (s, 1H) 23 466 [M + H]⁺ 0.84(C) 4.74-4.82 (m, 2H), 5.11 (dt, J = 7.1, 5.3 Hz, 1H), 5.50 (s, 2H),5.92 (d, J = 4.9 Hz, 1H), 7.33-7.40 (m, 4H), 7.68 (br s, 1H), 8.13 (brs, 1H), 8.76 (s, 1H)

Analytical HPLC methods

Method A

time Vol % water Flow (min) (incl. 0.1% TFA) Vol % ACN [mL/min] 0.00 991 1.6 0.02 99 1 1.6 1.00 0 100 1.6 1.10 0 100 1.6

Analytical column: XBridge BEH C18_2.1×30 mm, 1.7 μm; columntemperature: 60° C.

Method B

time Vol % water Flow (min) (incl. 0.1% TFA) Vol % ACN [mL/min] 0.00 973 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

Analytical column: Stable Bond (Agilent) 1.8 μm; 3.0×30 mm; columntemperature: 60° C.

Method C

time Vol % water Flow (min) (incl. 0.1% TFA) Vol % ACN [mL/min] 0.00 973 2.2 0.20 97 3 2.2 1.20 0 100 2.2 1.25 0 100 3.0 1.40 0 100 3.0

Analytical column: Sunfire (Waters) 2.5 μm; 3.0×30 mm; columntemperature: 60° C.

Method D

time Vol. % water Flow (min) (incl. 0.1% NH₄OH) Vol. % ACN [mL/min] 0.0095 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

Analytical column: XBridge C18_3.0×30 mm_2.5 μm (Waters); columntemperature: 60° C.

Method E

time Vol % water Flow (min) (incl. 0.1% TFA) Vol. % ACN [mL/min] 0.050.0 50.0 1.5 0.02 50.0 50.0 1.5 1.0 0.0 100.0 1.5 1.1 0.0 100.0 1.5

Analytical column: Sunfire (Waters) C18_2.1×30 mm_2.5 μm; columntemperature: 60° C.

Method F

time Vol. % water Flow (min) (incl. 0.1% NH₄OH) Vol. % ACN [mL/min] 0.0095 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5 1.60 95 5 1.5

Analytical column: XBridge C18 (Waters) 2.5 μm; 3.0×30 mm; columntemperature: 60° C.

Method G

time Vol. % water Flow (min) (incl. 0.1% TFA) Vol. % ACN [mL/min] 0.0099 1 1.6 0.02 99 1 1.6 1.0 0 100 1.6 1.1 0 100 1.6

Analytical column: Zorbax StableBond C18 (Agilent) 1.8 μm; 2.1×30 mm;column temperature: 60° C.

Method H

time Vol. % water Flow (min) (incl. 0.1% TFA) Vol. % ACN [mL/min] 0.0095 5 1.5 1.30 0 100 1.5 1.50 0 100 1.5

Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0×30 mm; columntemperature: 60° C.

1. A compound according to formula (I)

wherein A is selected from the group consisting of phenyl, naphthyl,thiophenyl, benzothiophenyl at and benzofuranyl, each optionallysubstituted with one or two members selected from the group consistingof H, F, Cl, Br, C₁₋₄-alkyl, F₁₋₃-fluoro-C₁₋₄-alkyl, —CN, —OCH₃,cyclopropyl, and cyclobutyl, or A is selected from

and R¹ is selected from H, C₁₋₄-alkyl, F₁₋₃-fluoro-C₁₋₄-alkyl,C₁₋₄-alkyl-OH or C₁₋₄-alkyl-CN; R² is selected from C₁₋₂-alkyl or Cl; orR¹ and R² are each CH₂ joined via a bond forming a 6-membered ring; andR³ is selected from H, or Ci-4-alkyl; or pharmaceutically acceptablesalt thereof.
 2. The compound of formula (I) according to claim 1,wherein A is selected from the group consisting of phenyl, thiophenyland benzofuranyl, each optionally substituted with one or two membersselected from the group consisting of H, F, Cl, Br, C₁₋₄-alkyl,F₁₋₃-fluoro-C₁₋₄-alkyl, CN, OCH₃, cyclopropyl, and cyclobutyl, or A isselected from

or pharmaceutically acceptable salt thereof.
 3. The compound of formula(I) according to claim 1, wherein A is selected from the groupconsisting of phenyl, thiophenyl and benzofuranyl, optionallysubstituted with one or two members selected from the group consistingof H, F, Br, Cl, and CH₃; or A is:

or pharmaceutically acceptable salt thereof.
 4. The compound of formula(I) according to claim 1, wherein A is selected from the groupconsisting of

or pharmaceutically acceptable salt thereof.
 5. The compound of formula(I) according to claim 1, wherein R¹ is selected from the groupconsisting of H and C₁₋₄-alkyl; or pharmaceutically acceptable saltthereof.
 6. The compound of formula (I) according to claim 1, wherein R²is CH₃ or Cl; or pharmaceutically acceptable salt thereof.
 7. Thecompound of formula (I) according to claim 1, wherein R³ is H or CH₃; orpharmaceutically acceptable salt thereof.
 8. The compound of formula (I)according to claim 1, selected from the group consisting of

or pharmaceutically acceptable salt thereof.
 9. The compound of formula(I) according to claim 1, selected from the group consisting of

or pharmaceutically acceptable salt thereof.
 10. A pharmaceuticallyacceptable salt of a compound according to claim
 1. 11. A pharmaceuticalcomposition comprising at least one compound of formula I according toclaim 1, or a pharmaceutically acceptable salt thereof, and one or morepharmaceutically acceptable excipients.
 12. A method for treating orpreventing an inflammatory airway disease, a fibrotic disease or coughcomprising administering to a patient having such disease or cough aneffective amount of a compound according to claim 1 or apharmaceutically acceptable salt thereof.
 13. A method according toclaim 12, wherein the patient has idiopathic pulmonary fibrosis (IPF) orcough.